Genetic encoding of a stable <i>O</i>-GlcNAc analogue

Attachment of N-acetylglucosamine to hydroxyl groups of serine and threonine residues of intracellular proteins is known as <i>O</i>-GlcNAcylation, an essential posttranslational event in mammals and most other metazoa, catalysed by a unique <i>O</i>-GlcNAc transferase (OGT) and removed by an <i>O</i>-GlcNAc hydrolase (OGA). Despite the existence of a single writer/eraser pair, proteomic studies have identified over 4000 O-GlcNAcylated proteins in the nucleus, cytoplasm and mitochondria of various organisms, ranging from <i>C. elegans </i>to human. However, the molecular and biological mechanistic consequences of site-specific <i>O</i>-GlcNAcylation have been understudied due to the lack of appropriate tools. The function of <i>O</i>-GlcNAc modification in the context of specific sites in vivo is usually examined by a loss-offunction Ser/Thr to Ala mutation. The only available tool to study gain-of-function <i>O</i>-GlcNAcylation in vivo is OGA inhibition, which causes global elevation of <i>O</i>-GlcNAcylation levels, complicating the dissection of site-specific modification. This thesis describes the development of approaches to study <i>O</i>-GlcNAcylation in a protein and site-specific manner. Due to the labile nature of <i>O</i>-GlcNAc and susceptibility to OGA hydrolysis, methods for site-targeted incorporation of its non-hydrolysable analogue were explored.

Name dc:type.qualificationname

Doctor of Philosophy

Level dc:type.qualificationlevel

Doctoral Thesis

Grantor dc:publisher.institution

University of Dundee

Year dc:date.issued

2018

Author dc:creator

• Gorelik, Andrii

Advisor dc:contributor.advisor

• van Aalten, Daan

Language dc:language

eng

Identifier

oai:discovery.dundee.ac.uk:studenttheses/ffddce97-0762-4c2b-96fb-d7a3666acffd

OAI identifier oai:identifier

oai:discovery.dundee.ac.uk:studenttheses/ffddce97-0762-4c2b-96fb-d7a3666acffd